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. 2021 Jan 6;7:1. doi: 10.1038/s41420-020-00384-x

Fig. 6. Defective myotube formation in Nogo-A-silenced C2C12 cells.

Fig. 6

Expression of Nogo-A in C2C12 myoblast was silenced using si-Nogo-A, and myoblast differentiation was induced by DM for 3 days. A, B qRT-PCR analysis of Nogo isoforms (A) and Myod, myogenin, and Myh2 (B) in si-scrambled- or si-Nogo-A-transfected C2C12 cells (n = 3/group). Mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001. C WB of indicated proteins in Nogo-A-silenced cells (n = 3/group). Molecular weights (kDa) are indicated. D Quantitative assessment of band intensities of WBs in C using the NIH ImageJ software. Mean ± SEM. **p < 0.01, ***p < 0.001. E IF staining of Nogo-A (green) with DAPI (blue) in differentiated si-scrambled- or si-Nogo-A-transfected C2C12 cells. Scale bar = 20 µm. F Fusion index of differentiated si-scrambled- or si-Nogo-A-transfected C2C12 cells. Fusion index was calculated as the percentage of total nuclei incorporated in myotubes. G Hematoxylin/eosin staining of differentiated si-scrambled- or si-Nogo-A-transfected C2C12 cells. Scale bar = 200 µm. H Quantitative assessment of myofiber diameter based on evaluation of 20 diameters in five fields per group) in L. Mean ± SEM. *p < 0.05.